猫疱疹病毒Ⅰ型的分离与鉴定

实验动物科学

猫疱疹病毒Ⅰ型的分离与鉴定

张硕;李纯玲;汪葆玥;刘金华;田克恭;

中国农业大学动物医学院;中国动物疫病预防控制中心;北京天之泰生物科技有限公司;

收稿日期:2010-04-28 修回日期:2010-04-28 出版日期:2010-04-28 发布日期:2010-04-28

Isolation and Identification of Feline Herpesvirus Type 1

ZHANG Shuo1,2,LI Chun-ling3,WANG Bao-yue2,LIU Jin-hua1,TIAN Ke-gong2

(1.College of Veterinary Medicine,China Agricultural University,Beijing 100094,China)(2.Chinese Center for Animal Disease Control and Prevention,Beijing 100094,China)(3.Tian Tech Biotechnology Co.Ltd,Beijing 100085,China

Received:2010-04-28 Revised:2010-04-28 Online:2010-04-28 Published:2010-04-28

摘要/Abstract

摘要： 目的从临床疑似猫病毒性鼻气管炎的病例中分离猫疱疹病毒Ⅰ型。方法用猫肾细胞(FK-81)对临床疑似猫病毒性鼻气管炎病猫的眼鼻分泌物进行分离培养,连续传4代,对获得的培养物进行了电子显微镜观察、血凝试验、病毒核酸型试验、理化特性试验以及PCR扩增和扩增产物基因测序鉴定。结果该病毒粒子呈球形,直径为160 nm左右,有囊膜。凝集猫红细胞,不凝集猪、兔、犬、小鼠和鸡的红细胞。病毒对乙醚、酸、热、胰蛋白酶敏感,其核酸型属DNA。PCR扩增为猫疱疹病毒Ⅰ型阳性,扩增产物基因序列与Nunberg等报道猫疱疹病毒Ⅰ型的基因序列同源性为100%(登录号:M26660)。结论在国内首次成功分离一株猫疱疹病毒Ⅰ型,其形态、血凝性、理化特性、基因序列等与国外文献报道一致,为猫病毒性鼻气管炎病原学、疫苗免疫、诊断及分子生物学研究奠定基础。

关键词: 猫疱疹病毒Ⅰ型, FK-81细胞

Abstract: Objective Isolate and cultivate Feline herpesvirus type 1 from clinical suspected cats.Methods The nasal and conjunctival secretion from the suspected cats were cultivated through 4 passages in FK-81 cells and collected,then identified by electron microscopy,hemocyte agglomeration test,physical and chemical character,the nucleic acid type assay,Polymerase Chain Reaction(PCR) amplification and genomic sequencsing.Results Virion was sphericity,and its diameter was approximately 160 nm with envelope.The isolated virus could agglutinate cat red blood cells,but it could not agglutinate red blood cells of pigs,rabbits,canines,mice and chooks.It was DNA virus and was sensitive to acid,heat,ether and trypsin.The isolated virus nucleotide was extracted and amplificated.it shared 100% homology with the sequence of the Feline herpesvirus type 1 in GenBank(ACCESSION:M26660).Conclusion The Feline herpesvirus type 1 was first isolated in China an dacoordinated with the foreign reports in morphology,hemagglutination,physical and chemical character and gene sequence.This finding will provide further research of Feline viral rhiontracheitis in pathology,molecular biology,diagnosis and vaccine.

Key words: Feline herpesvirus type 1, FK-81 cells, Isolation and Identification

张硕;李纯玲;汪葆玥;刘金华;田克恭;. 猫疱疹病毒Ⅰ型的分离与鉴定[J]. 实验动物科学.

ZHANG Shuo1,2,LI Chun-ling3,WANG Bao-yue2,LIU Jin-hua1,TIAN Ke-gong2. Isolation and Identification of Feline Herpesvirus Type 1[J]. laboratory animal science.